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FINAL YEAR PROJECT 2013

THE DYNAMICS OF ACETOWHITENING IN RABBIT CORNEAS AND TISSUE-ENGINEERED MODEL OF SQUAMOUS EPITHELIUM
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peggy khoo

on 12 February 2014

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Transcript of FINAL YEAR PROJECT 2013

PEGGY KHOO
THE DYNAMICS OF ACETOWHITENING IN RABBIT CORNEAS AND TISSUE-ENGINEERED MODEL OF SQUAMOUS EPITHELIUM
OVERVIEW
MATERIALS & METHODS
SUPERVISOR: DR STEPHEN MATCHER
FINAL YEAR PROJECT
MAY 2013
BACKGROUND
* Cervical cancer is the eleventh most common cancer in the UK.
* Acetowhitening phenomenon is used in conjunction with Colposcopy for the detection of pre-cancerous lesion.
* What is Acetowhitening effect?
* What is Colposcope?
* Limitation of Colposcopy - sensitive but not specific enough to confirm diagnosis.
BACKGROUND
* Optical Coherence Tomography (OCT) is an optical signal acquisition method based on low coherence interferometry technique.
* Non-invasive, high spatial resolution and enhanced depth penetration.
* Specific in the detection of micro-structural appearance in biological tissues.
AIM
Use OCT to image rabbit cornea and tissue-engineered model of squamous epithelium before and after topical addition of acetic and lactic acid
OBJECTIVES
To produce non-keratinised tissue-engineered model of squamous epithelium.
To characterise the dynamics of acetowhitening effect by quantifying the backscattering intensity at different depths of the multilayer tissues using Image J.
* Tissue-engineered model of squamous epithelium was produced by submerged culturing in Green's Media for 2 weeks.
PROTOCOL FOR OCT IMAGING
Samples were transferred into a well in petri-dish formed by blue tacks.
A reference OCT image was captured before the addition of acids.
Continuous OCT images were then captured upon application of 0.5mL acids in 0.5M, with a frequency of one image every 4 seconds for a total of 240 seconds.
The samples were immediately fixed in 3.7% formaldehyde and processed for immunohistochemistry.
RESULTS
Background
Aim & Objectives
Materials & Methods
Results
Conclusion & Further Work

* Non-inverted and inverted OCT were used in the study for rabbit cornea.
* Non-inverted OCT was used in the study for tissue-engineered skin composites.
TECHNICAL EXPERIMENTS
REFLECTIVITY STUDY ON MOLECULAR DIFFUSION
VIDEO SHOWING ACETOWHITENING PHENOMENON IN THE FIRST 100 SECONDS
0.5M OF ACETIC ACID
REFLECTIVITY CURVES ACROSS DIFFERENT DEPTHS OF RABBIT CORNEAS
BACKGROUND
* Why tissue-engineered model of squamous epithelium?
* Why rabbit corneas?
EPITHELIUM WHITENING AND PERMEABILITY OF ACIDS
RESULTS
RABBIT CORNEA
TISSUE-ENGINEERED SQUAMOUS EPITHELIUM
REFLECTIVITY CURVES ACROSS DIFFERENT DEPTHS OF MULTILAYERS SQUAMOUS EPITHELIUM
CONCLUSION
* First study on depth-resolved kinetics of acetowhitening.
* Quantitative results indicate a new aspect to the acetowhitening phenomenon.
* Contribute clinically towards the diagnosis of cervical cancer and hence, prevent the need of unnecessary biopsy and introducing long-term monitoring of the disease progression.
REFERENCES
[1] Massad, L.S et al (2009) Journal of Lower Genital Tract Disease 13: 137-144
[2] Maclean, Allan B (2004) Gynaecologic Oncology 95: 691–694
[3] Wu, Tao T et al (2005) Applied Optics 13: 4963–4973
[4] Balas, C et al (2008) Journal of selected topics in quantum electronics 14: 29-42
[5] Gallwas, Julia K.S et al (2011) Lasers in Surgery and Medicine 43: 206-212
[6] Kocsis E et al (1991) Journal of Structural Biology 107: 6-14
THANK YOU!!!
ANY QUESTIONS
FURTHER WORK
* Acetowhitening effect could be monitored in a prolonged period of time and across a wider depths.
* For the tissue-engineered skin, a different protocol could be introduced to reduce the artefacts caused by the change in optical path length.
0.5M OF LACTIC ACID
ACKNOWLEDGEMENT
I am truly and sincerely thankful to my project supervisor, Dr Stephen Matcher for providing me tremendous support and guidance to complete the project.
I would like to express my gratitude to Dr Shweta Mittar for her help in culturing the tissue-engineered model of squamous epithelium.
I would like to thank Dr Zeng Hai Lu for facilitating me in using Optical Coherence Tomography throughout the project.
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